title: Clinical validation of IFNγ/IL-10 and IFNγ/IL-2 FluoroSpot assays for the detection of Tr1 T cells and influenza vaccine monitoring in humans.
abstract: The type of T cell polarization and simultaneous production of multiple cytokines have been correlated with vaccine efficacy. ELISpot is a T cell detection technique optimized for the measurement of a secreted cytokine at the single cell level. The FluoroSpot assay differs from ELISpot by the use of multiple fluorescent-labeled anticytokine detection antibodies, allowing optimal measurement of multiple cytokines. In the present study, we show that an IFNγ/IL-10 FluoroSpot assay is more sensitive than flow cytometry to detect Tr1 regulatory T cells, an immunosuppressive T cell population characterized by the production of IL-10 and IFNγ. As many tolerogenic vaccines are designed to induce these Tr1 cells, this FluoroSpot test could represent a standard method for the detection of these cells in the future. The use of an IFNγ/IL-2 FluoroSpot assay during influenza vaccine monitoring showed that the influenza-specific IL-2-producing T-cell response was the dominant response both before and after vaccine administration. This study therefore questions the rationale of using the single-color IFNγ ELISpot as the standard technique to monitor vaccine-specific T-cell response. Using this same test, a trend was also observed between baseline levels of IFNγ T cell response and T cell vaccine response. In addition, a lower IFNγ+IL-2+ T-cell response after vaccine was observed in the group of patients treated with TNFα inhibitors (P=0.08). This study therefore supports the use of the FluoroSpot assay due to its robustness, versatility and the complementary information that it provides compared with ELISpot or flow cytometry to monitor vaccine-specific T-cell responses.
authors.affiliation: INSERM U970 ; Université Paris Descartes Sorbonne Paris-Cité; Paris, France; Hôpital Européen Georges Pompidou; Service d'Immunologie Biologique; Paris, France; CTL-Europe GmbH; Bonn, Germany.
authors.forename: Anne
authors.initials: A
authors.lastname: Chauvat
authors.affiliation: INSERM U970 ; Université Paris Descartes Sorbonne Paris-Cité; Paris, France; Hôpital Européen Georges Pompidou; Service d'Immunologie Biologique; Paris, France.
authors.forename: Nadine
authors.initials: N
authors.lastname: Benhamouda
authors.affiliation: INSERM U970 ; Université Paris Descartes Sorbonne Paris-Cité; Paris, France; Hôpital Européen Georges Pompidou; Service d'Immunologie Biologique; Paris, France.
authors.forename: Alain
authors.initials: A
authors.lastname: Gey
authors.affiliation: Departement de Biothérapie; Centre d'Investigation Clinique de biothérapie 1001; Groupe Hospitalier Pitié-Salpêtrière et Univ Pierre et Marie Curie Paris; Paris, France.
authors.forename: Francois M
authors.initials: FM
authors.lastname: Lemoine
authors.affiliation: Mabtech Ab; Nacka Strand, Sweden.
authors.forename: Staffan
authors.initials: S
authors.lastname: Paulie
authors.affiliation: Epidemiology, Information System, Modeling; UMR-S 707; University Paris 6-UPMC; Paris, France; Inserm U707; Paris, France; Public Health Unit; Saint-Antoine Hospital; Paris, France.
authors.forename: Fabrice
authors.initials: F
authors.lastname: Carrat
authors.affiliation: Antiviral Immunity; Biotherapy and Vaccine Unit; Infection and Epidemiology Department; Institut Pasteur; Paris, France.
authors.forename: Marie-Lise
authors.initials: ML
authors.lastname: Gougeon
authors.affiliation: Laboratoire de virologie; Hôpital Cochin; Assistance-Publique Hôpitaux de Paris (AP-HP); Paris, France.
authors.forename: Flore
authors.initials: F
authors.lastname: Rozenberg
authors.affiliation: Laboratoire de virologie; Hôpital Cochin; Assistance-Publique Hôpitaux de Paris (AP-HP); Paris, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Krivine
authors.affiliation: Departement de Biothérapie; Centre d'Investigation Clinique de biothérapie 1001; Groupe Hospitalier Pitié-Salpêtrière et Univ Pierre et Marie Curie Paris; Paris, France.
authors.forename: Mustapha
authors.initials: M
authors.lastname: Cherai
authors.affiliation: Cellular Technology Limited; Shaker Heights, OH USA.
authors.forename: Paul
authors.initials: P
authors.lastname: Lehmann
authors.affiliation: INSERM U970 ; Université Paris Descartes Sorbonne Paris-Cité; Paris, France.
authors.forename: Françoise
authors.initials: F
authors.lastname: Quintin-Colonna
authors.affiliation: Université Paris Descartes; Paris, France; Inserm; CIC BT505; Paris, France; Hôpital Cochin; AP-HP CIC de Vaccinologie Cochin-Pasteur; Paris, France.
authors.forename: Odile
authors.initials: O
authors.lastname: Launay
authors.affiliation: INSERM U970 ; Université Paris Descartes Sorbonne Paris-Cité; Paris, France; Hôpital Européen Georges Pompidou; Service d'Immunologie Biologique; Paris, France; Université Paris Descartes; Paris, France; Inserm; CIC BT505; Paris, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Tartour
mesh_terms: D058501:Enzyme-Linked Immunospot Assay / Q000379:methods; D005453:Fluorescence; D006801:Humans; D015151:Immunoblotting / Q000379:methods; D007252:Influenza Vaccines / Q000008:administration & dosage / Q000276:immunology*; D007251:Influenza, Human / Q000276:immunology / Q000517:prevention & control*; D007371:Interferon-gamma / Q000032:analysis*; D016753:Interleukin-10 / Q000032:analysis*; D007376:Interleukin-2 / Q000032:analysis*; D053244:Osteoprotegerin; D050378:T-Lymphocytes, Regulatory / Q000276:immunology*
chemical_list: C508609:IL10 protein, human; D007252:Influenza Vaccines; D007376:Interleukin-2; D053244:Osteoprotegerin; C506030:TNFRSF11B protein, human; D016753:Interleukin-10; D007371:Interferon-gamma
keywords: FluoroSpot; IL-10 suppressive T cells; T cell response; immunomonitoring; influenza vaccine